LOCUS       sequence-324506-        7853 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of Peptide coding gene (Binary vector).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7853)
  AUTHORS   Scheible W
  TITLE     Scheible Lab plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 7853)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "unpublished"
COMMENT     Sequence Label: LjUBQ:MtPIP1:T35s
FEATURES             Location/Qualifiers
     source          1..7853
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        45..224
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             246..1040
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERGRTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTQGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(300..319)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     910..929
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      1053..1305
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    1400..1424
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             1688..2317
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2746..3819
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGASSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      3885..4079
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        4362..4466
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4467..5261
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCAKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      5432..6020
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5921..5940
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6143..6167
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     polyA_signal    7624..7798
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
ORIGIN
        1 ccgccgatga cgcgggacaa gccgttttac gtttggaact gacagaaccg caacgttgaa
       61 ggagccactc agccgcgggt ttctggagtt taatgagcta agcacatacg tcagaaacca
      121 ttattgcgcg ttcaaaagtc gcctaaggtc actatcagct agcaaatatt tcttgtcaaa
      181 aatgctccac tgacgttcca taaattcccc tcggtatcca attagagtct catattcact
      241 ctcaaatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
      301 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
      361 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
      421 aactccaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
      481 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
      541 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
      601 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
      661 atcgcatcga gcgaggacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
      721 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcggatgc
      781 ccgacggcga ggatctcgtc gtgacccagg gcgatgcctg cttgccgaat atcatggtgg
      841 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
      901 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
      961 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     1021 ttcttgacga gttcttctga gcttgtcaag cagatcgttc aaacatttgg caataaagtt
     1081 tcttaagatt gaatcctgtt gccggtcttg cgatgattat catataattt ctgttgaatt
     1141 acgttaagca tgtaataatt aacatgtaat gcatgacgtt atttatgaga tgggttttta
     1201 tgattagagt cccgcaatta tacatttaat acgcgataga aaacaaaata tagcgcgcaa
     1261 actaggataa attatcgcgc gcggtgtcat ctatgttact agatcgacgc tggatccgaa
     1321 ttcggcaggg cccactgcat ccaccccagt acattaaaaa cgtccgcaat gtgttattaa
     1381 gttgtctaag cgtcaatttg tttacaccac aatatatcct gccaccagcc agccaacagc
     1441 tccccgaccg gcagctcggc acaaaatcac cactcgatac aggcagccca tcagccgtgc
     1501 ggctgcatga aatcctggcc ggtttgtctg atgccaagct ggcggcctgg ccggccagct
     1561 tggccgctga agaaaccgag cgccgccgtc taaaaaggtg atgtgtattt gagtaaaaca
     1621 gcttgcgtca tgcggtcgct gcgtatatga tgcgatgagt aaataaacaa atacgcaagg
     1681 ggaacgcatg aaggttatcg ctgtacttaa ccagaaaggc gggtcaggca agacgaccat
     1741 cgcaacccat ctagcccgcg ccctgcaact cgccggggcc gatgttctgt tagtcgattc
     1801 cgatccccag ggcagtgccc gcgattgggc ggccgtgcgg gaagatcaac cgctaaccgt
     1861 tgtcggcatc gaccgcccga cgattgaccg cgacgtgaag gccatcggcc ggcgcgactt
     1921 cgtagtgatc gacggagcgc cccaggcggc ggacttggct gtgtccgcga tcaaggcagc
     1981 cgacttcgtg ctgattccgg tgcagccaag cccttacgac atatgggcca ccgccgacct
     2041 ggtggagctg gttaagcagc gcattgaggt cacggatgga aggctacaag cggcctttgt
     2101 cgtgtcgcgg gcgatcaaag gcacgcgcat cggcggtgag gttgccgagg cgctggccgg
     2161 gtacgagctg cccattcttg agtcccgtat cacgcagcgc gtgagctacc caggcactgc
     2221 cgccgccggc acaaccgttc ttgaatcaga acccgagggc gacgctgccc gcgaggtcca
     2281 ggcgctggcc gctgaaatta aatcaaaact catttgagtt aatgaggtaa agagaaaatg
     2341 agcaaaagca caaacacgct aagtgccggc cgtccgagcg cacgcagcag caaggctgca
     2401 acgttggcca gcctggcaga cacgccagcc atgaagcggg tcaactttca gttgccggcg
     2461 gaggatcaca ccaagctgaa gatgtacgcg gtacgccaag gcaagaccat taccgagctg
     2521 ctatctgaat acatcgcgca gctaccagag taaatgagca aatgaataaa tgagtagatg
     2581 aattttagcg gctaaaggag gcggcatgga aaatcaagaa caaccaggca ccgacgccgt
     2641 ggaatgcccc atgtgtggag gaacgggcgg ttggccaggc gtaagcggct gggttgtctg
     2701 ccggccctgc aatggcactg gaacccccaa gcccgaggaa tcggcgtgag cggtcgcaaa
     2761 ccatccggcc cggtacaaat cggcgcggcg ctgggtgatg acctggtgga gaagttgaag
     2821 gccgcgcagg ccgcccagcg gcaacgcatc gaggcagaag cacgccccgg tgaatcgtgg
     2881 caagcggccg ctgatcgaat ccgcaaagaa tcccggcaac cgccggcagc cggtgcgtcg
     2941 tcgattagga agccgcccaa gggcgacgag caaccagatt ttttcgttcc gatgctctat
     3001 gacgtgggca cccgcgatag tcgcagcatc atggacgtgg ccgttttccg tctgtcgaag
     3061 cgtgaccgac gagctggcga ggtgatccgc tacgagcttc cagacgggca cgtagaggtt
     3121 tccgcagggc cggccggcat ggccagtgtg tgggattacg acctggtact gatggcggtt
     3181 tcccatctaa ccgaatccat gaaccgatac cgggaaggga agggagacaa gcccggccgc
     3241 gtgttccgtc cacacgttgc ggacgtactc aagttctgcc ggcgagccga tggcggaaag
     3301 cagaaagacg acctggtaga aacctgcatt cggttaaaca ccacgcacgt tgccatgcag
     3361 cgtacgaaga aggccaagaa cggccgcctg gtgacggtat ccgagggtga agccttgatt
     3421 agccgctaca agatcgtaaa gagcgaaacc gggcggccgg agtacatcga gatcgagcta
     3481 gctgattgga tgtaccgcga gatcacagaa ggcaagaacc cggacgtgct gacggttcac
     3541 cccgattact ttttgatcga tcccggcatc ggccgttttc tctaccgcct ggcacgccgc
     3601 gccgcaggca aggcagaagc cagatggttg ttcaagacga tctacgaacg cagtggcagc
     3661 gccggagagt tcaagaagtt ctgtttcacc gtgcgcaagc tgatcgggtc aaatgacctg
     3721 ccggagtacg atttgaagga ggaggcgggg caggctggcc cgatcctagt catgcgctac
     3781 cgcaacctga tcgagggcga agcatccgcc ggttcctaat gtacggagca gatgctaggg
     3841 caaattgccc tagcagggga aaaaggtcga aaaagcttct ttcctgtgga tagcacgtac
     3901 attgggaacc caaagccgta cattgggaac cggaacccgt acattgggaa cccaaagccg
     3961 tacattggga accggtcaca catgtaagtg actgatataa aagagaaaaa aggcgatttt
     4021 tccgcctaaa actctttaaa acttattaaa actcttaaaa cccgcctggc ctgtgcataa
     4081 ctgtctggcc agcgcacagc cgaagagctg caaaaagcgc ctacccttcg gtcgctgcgc
     4141 tccctacgcc ccgccgcttc gcgtcggcct atcgcggccg ctggccgctc aaaaatggct
     4201 ggcctacggc caggcaatct accagggcgc ggacaagccg cgccgtcgcc actcgaccgc
     4261 cggcgcccac atcaaggcac cgagctccca tccccccagg ggctgcgccc ctcggccgcg
     4321 aacggcctca ccccaaaaat ggcagcgctg gccaattcgt gcgcggaacc cctatttgtt
     4381 tatttttcta aatacattca aatatgtatc cgctcatgag acaataacct tgataaatgc
     4441 ttcaataata ttgaaaaagg aagagtatgg ctaaaatgag aatatcaccg gaattgaaaa
     4501 aactgatcga aaaataccgc tgcgcaaaag atacggaagg aatgtctcct gctaaggtat
     4561 ataagctggt gggagaaaat gaaaacctat atttaaaaat gacggacagc cggtataaag
     4621 ggaccaccta tgatgtggaa cgggaaaagg acatgatgct atggctggaa ggaaagctgc
     4681 ctgttccaaa ggtcctgcac tttgaacggc atgatggctg gagcaatctg ctcatgagtg
     4741 aggccgatgg cgtcctttgc tcggaagagt atgaagatga acaaagccct gaaaagatta
     4801 tcgagctgta tgcggagtgc atcaggctct ttcactccat cgacatatcg gattgtccct
     4861 atacgaatag cttagacagc cgcttagccg aattggatta cttactgaat aacgatctgg
     4921 ccgatgtgga ttgcgaaaac tgggaagagg acactccatt taaagatccg cgcgagctgt
     4981 atgatttttt aaagacggaa aagcccgaag aggaacttgt cttttcccac ggcgacctgg
     5041 gagacagcaa catctttgtg aaagatggca aagtaagtgg ctttattgat cttgggagaa
     5101 gcggcagggc ggacaagtgg tatgacattg ccttctgcgt ccggtcgatc agggaggata
     5161 tcggggaaga acagtatgtc gagctatttt ttgacttact ggggatcaag cctgattggg
     5221 agaaaataaa atattatatt ttactggatg aattgtttta gctgtcagac caagtttact
     5281 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5341 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5401 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5461 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5521 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     5581 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5641 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5701 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5761 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5821 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5881 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     5941 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6001 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctgtggttgg
     6061 cacatacaaa tggacgaacg gataaacctt ttcacgccct tttaaatatc cgattattct
     6121 aataaacgct cttttctctt aggtttaccc gccaatatat cctgtcaaac actgatagtt
     6181 taaactccct cggtcacatg tgcatcctct agtggaggga gagaggattt tgaggaaata
     6241 attaattgaa ttacttgtat tattgataaa gtaatttaga taagttgtta gtacaactta
     6301 ttgcaactat ggatagacaa aaatcactta ttttaagggg agaagtatat aacaacttat
     6361 tataaaattt ctgagcagcc ggcctcctcc aatcaatcat agagagtgga gcccattctg
     6421 gaaaaccaag gaacccccaa cttgcactgg tgcggtggcc caattcaaaa aaaacggggc
     6481 ccagcaaagt aacgccgttg taacgattta tcaatccaaa ctcaaaaggc ggcgagatgc
     6541 gtttaactcc gtgaaattaa caaaccgcca acaacttgca atttgcaact accgtttcca
     6601 agaagaactc aaccacacaa cgtatcctat cccaaaccac acgcaactag tgacgcgtca
     6661 taaggacacg tgtcacaatt tgactggtta ataatttcac cgcttttgct ataaattacc
     6721 tccaatcccc ttagcttctt cacaattcag ttcccaaccc taacaattct tgttcatatc
     6781 gcttctctct actttcaagg tatgatccaa tttctctctt ctttctctgt aatcctttcg
     6841 ttgagttttg tttccgatca atcataggta gttttcttgt ttcgaagcat gagatctagg
     6901 aattttttgt gattttccaa aattgagatc ggtttgaaat tgaattttac agcttgaatc
     6961 tcagatcttg ttttatcaat gttttcgatg gcttgcgatg tagatctatg ataattgtgg
     7021 ttcagttttg ttaggaatcg atttcggttt agcaattgca gattaattag ggtttccaat
     7081 tgaattcttc agatccgtta tggaattatg tcaaataatt tattcaaatt ggaaattatt
     7141 gttagatcca ctcttaatct gtttgatcca agcttcaatt agggttttca cttgtttcaa
     7201 tttcttgtta tggattctga tttatctgtt gatgttagat ccactcttaa tctgtttgat
     7261 ccaagcgtta attagggttt tcacttgttt caatttcgtg tgttggattc tgatttatct
     7321 gttgttgatg tgattacaga atgagttatg taaaggcatg tacccttttg aatttgatca
     7381 caattgttat gatctgtttg gctattttga tgagtgaaag tcaaagtagt aaggtggagg
     7441 caagtagggt tttaaggaat tcacaagagt ttgcatttgc aagagctaat caccttcaaa
     7501 catcagcata tgaacaggcc aagaacacaa tgacaatttg gcttcaacgt ttgccttctg
     7561 gaccaagtcc taaaggccct ggtcattgag cttctctagc tagagtcgat cgacaagctc
     7621 gagtttctcc ataataatgt gtgagtagtt cccagataag ggaattaggg ttcctatagg
     7681 gtttcgctca tgtgttgagc atataagaaa cccttagtat gtatttgtat ttgtaaaata
     7741 cttctatcaa taaaatttct aattcctaaa accaaaatcc agtactaaaa tccagatcgc
     7801 taagcttgaa ttcttgcgga gagcggagaa ttaagggagt cacgttatga ccc
//
